Figure 5.
Single cut and read-out approaches. Dephosphorylated DNA is split into two separate reactions. crRNA guides are designed in a strand-directed manner with separate guide pools prepared for guides cutting upstream versus downstream from each target. Upstream and downstream guide pools are then used to cleave dephosphorylated DNA in separate reactions. After Cas9 cleavage, both reactions are pooled together, and sequencing adapters are ligated. The prepared library is loaded on nanopore flowcells and sequenced. (Created with BioRender; https://www.biorender.com/)
